Penicillins

ABSTRACT

New penicillins and their salts and esters particularly active against Gram-negative organisms such as Pseudomonas spp., and their preparation and administration. The penicillins are active against clinically important organisms against which well-known broad spectrum penicillins are inactive and may generally be designated as aminoacyldipeptide penicillins of unusual structure and properties.

CROSS-REFERENCE

This is a division of Ser. No. 466,814 filed May 3, 1974.

This invention relates to penicillins which have, in general, a broadspectrum of antibacterial activity, being active against many species ofGram-positive and Gram-negative bacteria. They are thus useful astherapeutic (and, to a lesser extent, prophylactic) agents in animals,including man and poultry. The invention further relates to methods forthe preparation of these penicillins and to their use in therapy.

Although there are now available a number of semi-synthetic penicillinshaving what is known as broad-spectrum activity, no single penicillin isyet available which has a clinically useful level of antibacterialactivity against all the pathogenic organisms encountered in clinicalpractice. The search thus continues for broad-spectrum penicillins whichhave advantages, either in improved antibacterial effectiveness or widerspectrum of activity, over the available penicillins.

According to the present invention there is provided a penicillin offormula (I) or a pharmaceutically acceptable salt or ester thereof:##SPC1##

Wherein

R is phenyl, phenyl substituted by one or more functional groupsselected from hydroxy, halogen, nitro, alkoxy containing from 1 to 3carbon atoms, and amino groups, 2- or 3- thienyl, cycloalkyl having from3 to 7 carbon atoms, cycloalkenyl having from 5 to 7 carbon atoms oralkyl having from 1 to 4 carbon atoms;

R³ is hydrogen or an alkyl group having from 1 to 3 carbon atoms;

R¹ is hydrogen or an organic radical containing up to 20 carbon atoms;

R² is a group of formula (II) or (III): ##EQU1## wherein R⁵ is amino,mono- or di-alkylamino wherein the alkyl groups contain from 1 to 4carbon atoms, cyclohexylamino, hydrogen, alkyl having from 1 to 4 carbonatoms, or phenyl and R⁶ is amino or mono- or di- alkylamino wherein thealkyl groups contain from 1 to 4 carbon atoms, or cyclohexylamino.

The group R may be, for example, phenyl, 4-hydroxyphenyl,3-chloro-4-hydroxyphenyl, 4-nitrophenyl, 4-chlorophenyl, 4-fluorophenyl,4-methoxy-phenyl, 4-aminophenyl, 2-thienyl, 3-thienyl, cyclopropyl,cyclohexyl, cyclohexa-1,4-dienyl, iso-propyl or methyl.

The group R¹ may, for example, be hydrogen, phenyl, 4-hydroxyphenyl,4-nitrophenyl, 4-chlorophenyl, 4-fluorophenyl, 4-methoxyphenyl,4-aminophenyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,tertbutyl, methoxy, ethoxy, n-propoxy, isopropoxy, methylene, ethylene,ethylthio, n-propoxy-methyl, carbamyl, carbamylmethyl, acetoxy, phenoxy,benzyloxy, 2-thienyl, 3-thienyl, indol-3-yl, 1H-imidazol-5-yl,cyclohexa-1,4-dienyl, cyclopropyl or cyclohexyl.

The group R⁵ may, for example, be amino, methylamino, n-butylamino,tert-butylamino, cyclohexylamino, hydrogen, methyl, ethyl, n- orisopropyl, n-, sec, or tert-butyl, or phenyl.

The group R⁶ may, for example, be amino, methylamino, dimethylamino,ethylamino, diethylamino, n-propylamino, isopropylamino,tert-butylamino, n-butylamino or cyclohexylamino.

Preferably R is phenyl, 4-hydroxyphenyl, or 3-thienyl.

Preferably R¹ is phenyl, 4-hydroxyphenyl, 4-chlorophenyl,4-fluorophenyl, 4-nitrophenyl, 3-indolyl, or methylthiomethyl.

Preferably R³ is hydrogen.

Preferably R⁵ is amino or hydrogen.

Preferably R⁶ is amino.

Preferably the carbon atom to which the group R in formula (I) isattached is in the D configuration.

Preferably the carbon atom to which the group R² in formula (I) isattached is in the D configuration.

Examples of suitable salts of compounds (I) include the sodium,potassium, calcium, magnesium or aluminium salts, and ammonium orsubstituted ammonium salts, for example those with trialkylamines suchas triethylamine, procaine, dibenzylamine, triethanolamine,1-ephenamine, ethylpiperidine, and other amines which have been used toform salts with benzylpenicillins. In the case of compounds (I) whichcontain a basic nitrogen site in the side chain, acid addition salts mayalso be formed. Such salts include, for example, inorganic salts such asthe sulphate, nitrate, phosphate, borate, thiocyanate, and hydrohalides,e.g. hydrochloride, hydrobromide and hydroiodide, and organic salts suchas the acetate, oxalate, tartrate, malate, citrate, succinate, benzoate,ascorbate and methanesulphonate.

Examples of suitable pharmaceutically acceptable esters include thosewhich break down readily in the human body to leave the parent acid,e.g., acyloxyalkyl esters such as acetoxymethyl, pivaloyloxymethyl,α-acetoxyethyl, α-acetoxybenzyl and α-pivaloyloxymethyl esters, andalkoxycarbonylalkyl esters such as methoxy carbonyloxymethyl esters.Other suitable esters of this readily hydrolysable type include lactone,thiolactone, and dithiolactone esters (i.e. compounds of formula (I)wherein the 3-carboxy group is esterified to produce a grouping offormula: ##EQU2## wherein X¹ and Y¹ are oxygen or sulphur and Z¹ is adivalent hydrocarbon group), especially the phthalidyl and substitutedphthalidyl esters e.g. 5,6-dimethoxyphthalidyl ester.

The compounds of this invention may be prepared by reacting6-aminopenicillanic acid or a salt, ester or silyl derivative thereofwith an N-acylating derivative of an acid of formula (IV) ##EQU3## inwhich any reactive substituents may be blocked, wherein R, R¹, R² andR³, are as defined in formula (I) and then, if necessary, carrying outone or more of the following steps (i) removing any silyl groups byhydrolysis or alcoholysis, (ii) converting an ester compound to a freeacid or salt (iii) converting a salt to a free acid or a free acid to asalt (iv) removing any blocking groups to release the desired functionalsubstituent (v) converting a free acid compound to an ester compound.

By the term "silyl derivative" used in connection with6-aminopenicillanic acid (6-APA) we mean the product of the reactionbetween 6-APA and a silylating agent such as a halotrialkylsilane,halodialkylsilane, a halotrialkoxysilane, a dihalodialkoxysilane or acorresponding aryl or aralkyl silane and compounds such ashexamethyldisilazane. In general, halotrialkylsilanes are preferred,especially trimethylchorosilane.

A reactive N-acylating derivative of the acid (IV) is employed in theabove process. The choice of reactive derivative will of course ofinfluenced by the chemical nature of the substituents in the acid. Thus,when the acid contains only acid stable groups, an acid halide is asuitable N-acylating derivative, preferably the acid chloride.

Such reagents would, however, be avoided when an acid labile group waspresent in the acid (IV). In such cases a suitable N-acylatingderivative is a mixed anhydride. For this purpose particularlyconvenient mixed anhydrides are the alkoxyformic anhydrides.

However, with both the acid chloride and mixed anhydride N-acylatingagents we have found that some racemisation may take place. To minimisesuch unwanted racemisation, we prefer to use an activated ester as theN-acylating agent. Such activated esters, for example the ester formedwith 1-hydroxybenzotriazole or, preferably, N-hydroxysuccinimide, may beprepared in situ by the reaction of the acid with the appropriatehydroxy compound in the presence of a carbodiimide, preferablydicyclohexylcarbodiimide.

Other reactive N-acylating derivatives of the acid (II) include thereactive intermediate formed by reaction in situ with a carbodiimide orcarbonyldiimidazole, but the literature on the preparation ofsemisynthetic penicillins contains examples of other reactiveN-acylating derivatives of acids suitable for coupling to 6-APA.

It will be understood, of course, that where a free acid of type (I) ora salt thereof is desired, it may be convenient to carry out theacylation reaction using an ester of 6-APA, and then to remove the estergroup. Vice versa, if an ester is required, it may be convenient tocarry out the acylation reaction using 6-APA or a salt thereof andthereafter to esterify the free acid.

In the above process, if it is necessary to block any reactivesubstituents in the acid (IV), conventional chemical blocking groups areknown. Thus, if desired, any free amino groups may be blocked byconversion to benzyloxycarbonylamino groups, or the amino group may beblocked as the nitro group which is later converted to the amino group.

The compounds of this invention may also be prepared by a process whichcomprises reacting a compound of formula (V) or a salt, ester or silylderivative thereof. ##SPC2##

wherein R is as defined in formula (I) and in which any reactivesubstituents may be blocked, with an N-acylating derivative of an acidof formula (VI) ##EQU4## wherein R¹, R² and R³ are as defined in formula(I), and if necessary, carrying out one or more of the following steps(i) removing any silyl groups by hydrolysis or alcoholysis, (ii)converting an ester compound to a free acid or salt thereof (iii)converting a salt to a free acid or a free acid to a salt (iv) removingany blocking groups to release the desired functional substituents (v)converting a free acid compound to an ester compound.

The remarks made earlier with respect to silyl derivatives, N-acylatingderivatives, and blocking groups, also apply to this process.

The compounds of this invention wherein R¹ is a group of formula (II)may also be prepared by reacting a compound of formula (VII) or a salt,ester or silyl derivative thereof: ##SPC3##

wherein R, R¹, R³ and R⁴ are as defined in formula (I) and wherein anyreactive substituents may be blocked, with cyanate ion, a C₁ ₋₄ alkylisocyanate, cyclohexyl isocyanate, a formylating agent or an N-acylatingderivative of an acid R⁶ COOH wherein R⁶ is phenyl or an alkyl grouphaving from 1 to 4 carbon atoms, followed, if necessary, by one or moreof the following steps (i) removing any silyl groups by alcoholysis orhydrolysis, (ii) converting an ester compound to a free acid or saltthereof, (iii) converting a salt to a free acid or a free acid to a salt(iv) removing any blocking groups to release the desired functionalsubstituent, (v) converting a free acid compound to an ester compound.

It will be noted that the above process essentially consists ingenerating the desired group R² from the free amino group in compound(VII). The reaction of amino compounds with cyanate ion and isocyanatesto produce ureas and substituted ureas is well known. Likewise theformylation of amino compounds (e.g., using formic acid and aceticanhydride) is well known. Similarly, the acylation of amino compounds isextremely well known, and suitable N-acylating derivatives of acids havebeen discussed hereinbefore.

The compounds of this invention are broad spectrum penicillins, i.e.penicillins which not only have activity against Gram-positive bacteria,but also against a number of clinically important Gram-negativeorganisms. The preferred compounds of this invention are active againstsuch important organisms as Pseudomonas spp. against which the most wellknown broad-spectrum penicillin(6[(D)α-aminophenylacetamido]penicillanic acid . . . ampicillin) isnormally inactive. Moreover the preferred compounds of this inventionare about as active as 6[(D)α-carboxy-3-thienylacetamido] penicillanicacid against Pseudomonas spp., this latter compound being the mostactive of the known penicillins against those organisms. Several of thepreferred compounds of this invention have minimum inhibitoryconcentrations of from 5 - 12.5 μg/ml against some β-lactamase producingstrains of staphylococci, against which the majority of known broadspectrum penicillins are only marginally effective. The preferredcompounds of this invention are not greatly serum-bound, and are notmarkedly inactivated by serum.

The penicillins of this invention show the characteristic lack oftoxicity of penicillins generally. They may be administered byparenteral injection. The daily dose will depend on the identity of thepencillin and severity of infection. With the preferred compounds ofthis invention, a suitable average daily dose for an adult would be inthe range of 100mg to 5000mg. An average single dose for an adult wouldbe from 20 mg to 500 mg.

The following Examples illustrate the preparation of some of thecompounds of this invention: ##SPC4##

In the following Examples amoxycillin is the approved name for6[D-α-amino-p-hydroxyphenylacetamido]penicillanic acid and ampicillin isthe approved name for 6[D-α-aminophenylacetamido] penicillanic acid.Epicillin is the approved name for [D-α-amino-cyclohexa-1,4,-dienylacetamido]penicillanic acid. All temperatures arein °C. All biochromatograms were run in butanol/ethanol/water. Allcompounds were made by one of the following generally applicablemethods.

The majority of the starting materials used in the following Examplesare known. However, the following literature references describegenerally applicable methods which may be used to prepare the startingmaterials:

UREIDO-ACIDS

DAKIN: Amer.Chem. J. 44 54

ANDREASCH: Monats. 23. 805

NEVILLE, M^(c) GEE: Can.J.Chem. 41, 2123-9 (1963)

WIELAND: Bio.Z. 38, 389, Ann.3.

DAVIS, BLANCHARD: J.Amer.Chem.Soc. 51, 1797

LEUTHARDT,BRUNNER: Helv.Chim.Acta. 30, 964-5 (1947)

SUBSTITUTED UREIDO-ACIDS

BALL, SKINNER, SHIVE: Texas Rept. Biol.Med. 21(2) 188-75 (1963)

BRITISH PATENTS 1301961/2.

GUANIDINO-ACIDS

KAPFHAMMER, MILLER: Z. Physiol,Chem. 225, 1-12, (1934)

RADKA PANT: Ibid 335, 272-4 (1964)

FRAMM, KAPELLER: Ann. (1925) 442, 144

HABEL: Can.J.Biochem. Physiol. 38, 493 (1960)

RAMSAY: Ber. 41, 4390

FORMAMIDO-ACIDS

SHEEHAN, YOUNG: J.Amer.Chem.Soc. (1958), 80, 1154

METHOD A

A solution of the guanidino-acid, hydrochloride (5m. mole) in drydimethylformamide (5 ml) was added over 10 mins. to a stirred solutionof phthalid-3-yl D-α-aminophenyl-acetamidopenicillanate (5 m. mole) andN,N¹ -dicyclohexylcarbodi-imide (5.8 m. mole) at 0°C in dry methylenedichloride.

After stirring at 0°C for 30 mins. and 11/2 hours at ambienttemperatures, the mixture was cooled to -10°C and the dicyclohexylurearemoved by filtration.

The solution was washed with dilute hydrochloric acid (pH 1.5), water,and brine and the dried solution concentrated to low volume in vacuo toinduce crystallisation. The filtered solid was dried under vacuum overphosphorus pentoxide.

METHOD B

Ureido- (or substituted-ureido-) acid (0.01 mole) in dry acetone (60 ml)at -10°C was treated with triethylamine (ca. 0.015 mole) andiso-butylchloroformate (0.01 mole) and stirred at -10°C for not morethan 30 mins. D-α-aminophenylacetamidopenicillanic acid, trihydrate(0.01 mole) in water (60 ml) was treated with triethylamine to give aclear solution (pH 8.4). Acetone (60 ml) was added and the solutioncooled to 0°C.

The mixed anhydride solution cooled to -40°C was filtered through Celiteinto the stirred penicillin solution and the mixture allowed to warmslowly to room temperature over 20 mins.

The acetone was evaporated in vacuo and the aqueous residue washed wellwith ether and then acidified to pH2 under a layer of ethyl acetate with5N hydrochloric acid.

The product was obtained either as the free acid by filtration of theaqueous/ethyl acetate mixture or by precipitation from the ethyl acetatesolution with potassium or sodium 2-ethylhexoate to give thecorresponding alkali-metal salt.

Method Bi. As B, but using N-methylmorpholine instead of triethylaminein the preparation of the mixed anhydride.

Method Bii. As B, but usingD-α-amino(p-hydroxyphenyl)acetamidopenicillanic acid, trihydrate insteadof D-α-aminophenylacetamidopenicillanic acid, trihydrate.

Method Biii. As (Bii) but using N-methylmorpholine instead oftriethylamine in the preparation of the mixed anhydride.

Method Biv. As (Bi), but usingD-α-amino-(3-thienyl)-acetamidopenicillanic acid instead ofD-α-aminophenylacetamidopenicillanic acid, trihydrate.

Method Bv. As (Bi), but usingD-α-amino-(1,4-cyclohexadienyl)acetamidopenicillanic acid instead ofD-α-aminophenylacetamidopenicillanic acid, trihydrate.

Method Bvi. As (Bi), but using ethylchloroformate instead of iso-butylchloroformate and D-α-aminocyclopropylacetamidopenicillanic acid,trihydrate.

Method Bvii. As (Bi), but using D-α-aminovaleramidopenicillanic acidinstead of D-α-aminophenylacetamidopenicillanic acid, trihydrate.

Method Bviii. As (Bi), but using ethylchloroformate instead of isobutylchloroformate, and D-α-amino-(2-thienyl)-acetamidopenicillanic acidinstead of D-α-aminophenylacetamidopenicillanic acid, trihydrate.

Method Bix. As (Bi), but usingD-α-amino-β-phenylpropionamidopenicillanic acid instead ofD-α-aminophenylacetamidopenicillanic acid.

(No Method C)

Method D

Amino-penicillin (5 m.moles) in dry dimethylformamide (100ml) wastreated with triethylamine (12 m.moles) and stirred to give a clearsolution. Sulphur trioxide-triethylamine complex (6 m.moles) was addedportionwise over 5 mins. at room temperature and stirred for 1 hour. Asolution of potassium 2-ethyl hexoate (ca 15 m.moles) in dry acetone(150ml) was added and a white solid separated.

After further dilution with acetone (200ml) the solid was filtered,washed with acetone and then stirred in dry ether for 20 mins. to removeany residual dimethylformamide. The solid was filtered and dried invacuo.

METHOD E

Anhydrous D-α-aminophenylacetamidopenicillanic acid (5m. mole) in drymethylene dichloride (50 ml) was treated with triethylamine (˜10 m.mole) to give a clear solution. Trimethylsilyl chloride (10 m. mole) wasadded and the mixture refluxed under nitrogen for 1 hour, then cooled to0°C.

α-Guanidino-acid (5 m. mole) was dissolved in dry dimethylformamide (5ml) and dry dimethylformamide (5 ml.) and dry methylene dichloride (50ml) added, cooled to 0°C and stirred for 5 mins. withdicyclohexyl-carbodi-imide (5.5 m. mole). The bis-trimethylsilylatedpenicillin was added and stirred at 0°C for 1 hour. The mixture was thencooled to -20°C and the dicyclohexylurea removed by filtration. Thefiltrate was evaporated to dryness in vacuo and the residue dissolved inacetone (20 ml)/water (20 ml) and the pH adjusted to 2.5 with 5Nhydrochloric acid. After stirring at pH 2.5 for 25 mins. the acetone wasremoved in vacuo and any solid filtered off. The residual aqueoussolution was freeze dried and the resultant solid treated with water atpH 2. The product was filtered and dried.

METHOD F

Dicyclohexylcarbodi-imide (5.5 m. mole) was added to a stirred solutionof N-substituted-amino acid (5 m. mole) in dry acetone (20 ml) at 0°C.The mixture was stirred for 15 mins. at 0°-5°C and then left in therefrigerator overnight.

D-α-Aminophenylacetamidopenicillanic acid, trihydrate (5m. mole) wasdissolved in acetone (10 ml)/water (10 ml) with triethylamine (0.7 ml)and the hydroxysuccinimide ester filtered in, through Celite. Afterstirring for 45 mins. the acetone was removed in vacuo, leaving agelatinous mass. Acidification with 5N hydrochloric acid in aqueousethyl acetate gave the product as the free acid (sometimes only afterconcentration of the ethyl acetate layer and treatment with ether) or asthe salt by treatment of the washed and dried ethyl acetate layer withsodium or potassium 2-ethyl hexoate.

Method Fi. As F, but the hydroxysuccinimide ester formed in drydimethylformamide (or dimethylformamide diluted with acetone).

Method Fii. As F, but the hydroxysuccinimide ester formed in dry1,2-dimethoxyethane.

Method Fiii. As (Fi), but the penicillin dissolved inacetone/chloroform, the product coming out of solution as the aminesalt.

EXAMPLE 1D-α-[D-β-(p-Hydroxyphenyl)-α-ureidopropionamido]phenylacetamidopenicillanic acid

(R = Ph; R¹ = --p--HO--PhCH₂ ; R³ = H; R² = NHCONH₂ ; M = H; α¹ = D).Prepared by method (Bi), from D-β-(-p-Hydroxyphenyl)-α-ureidopropionicacid.

Yield: 68%

νmax (KBr): 3550, 1770, 1650, 1515, 1230 and 700cm⁻ ¹

δ[(CD₃)₂ SO]: 1.44 (3H. s.gem methyl); 1.57 (3H. s. gem methyl); ˜2.8(2H. m. -CH₂ CH<); 4.29 (1H. s. C-3 proton); ˜4.5 (1H. m. -CH₂ CH<);5.35 - 5.87 (5H. m. β-lactams, Ph CH-; NHCONH₂ *) 6.27 (1H. d. NHCONH₂*); ##SPC5##

7.30 (5h. broad s. Ph CH<); 8.47 (1H. m. --CONH*); 9.12 (1H. m.--CONH*--).

NH₂ OH Assay: 101%

Biochromatography: 1 zone at R_(f) 0.32

EXAMPLE 2D-α-[D-β-(-p-Hydroxyphenyl)-α-ureidopropionamido]-(-p-hydroxyphenyl)acetamidopenicillanic acid.

(R = -p-HO-Ph; R¹ = -p-HO-PhCH₂ ; R³ = H; R² = NHCONH₂ ; M=H; α¹ =D).Prepared by method (Bi), from D-β-(-p-hydroxyphenyl)β-ureidopropionicacid and amoxycillin.

Yield: 74%

νmax (KBr); 3350 (broad), 1770, 1650, 1515, 1230 and 840 cm⁻ ¹.

δ[(CD₃)₂ SO]: 1.42 (3H. s gem methyl); 1.57 (3H. s. gem methyl); ˜2.8(2H. m. --CH₂ CH<); 4.3 (1H.s. C-3 proton); ˜4.4 (1H. m. CH₂ CH<); 5.3 -5.85 (5H. m. β-lactams, Ph CH-- ; NHCONH₂ *); 6.25 - 7.30 (8H. m.aromatics protons); 8.47 (1H. m. CONH*--); 9.12 (1H. m. --CONH*--).

NH₂ OH assay: 94%

Biochromatography: 1 zone at R_(f) = 0.18.

EXAMPLE 3 D-α-[D-β-(p-Hydroxylphenyl)-α-ureidopropionamido](3-thienyl)acetamido penicillanic acid. ##SPC6##

R¹ = p--HO--PhCH₂ ; R³ = H; R² = NHCONH₂ ; M=H; α¹ =D). Prepared bymethod (Bii), from D-β-(p-hydroxyphenyl)-α-ureidopropionic acid andα-amino-(3-thienyl) acetamido penicillanic acid.

Yield: 72%

νmax (KBr): 3350 (broad); 1770, 1650, 1515, 1230, 845 and 780cm⁻ ¹.

δ[(CD₃)₂ SO]: 1.45 (3H. s. gem methyl); 1.58 (3H s. gem methyl); ˜2.8(2H. m. --CH₂ CH<); 4.3 (1H.s . C-3 proton), ˜4.4 (1H. m. --CH₂ CH<);5.35 - 5.90 (5H. m. β-lactams, Ph --CH -- ; NHCONH₂ *); 6.25 (1H. m. NHCONH₂); 6.5 - 7.6 (7H. m. aromatics); 8.5 (1H. d. CONH*); 9.1 (1H.d.CONH*).

NH₂ OH assay: = 96%

Biochromatography: 1 zone at R_(f) = 0.31

EXAMPLE 4D-α-[DL-β-(-p-Nitrophenyl)-α-ureidopropionamido]-(-p-hydroxyphenyl)acetamidopenicillanicacid.

(R = p-HO--Ph; R¹ = p-NO₂ --PhCH₂ ; R³ = H; R² = NHCONH₂ ; M=H; α¹ =DL). Prepared by method (Bii), fromDL-β-(-p-Nitrophenyl)-α-ureidopropionic acid and amoxycillin.

Yield: 60%

νmax (KBr): 3350, 1770, 1650, 1514 and 1230 cm⁻ ¹.

δ[(CD₃)₂ SO]: 1.42 (3H. s. gem methyl); 1.55 (3H. s. gem methyl); 3.0(2H. m. CH₂ CH<); 4.05 (1H. s. C-3 proton); ˜4.60 (1H. m --CH₂ CH<);5.25 - 5.80 (5H. m. β-lactams, ##SPC7##NHCONH₂ * ); 6.2 (1H. m.NH*CONH₂); 6.5 - 8.2 (8H. m. aromatics); 8.50, 9.00 (2 x 1H.m. CONH*).

NH₂ OH assay: 86.5%

Biochromatography: 1 zone at R_(f) = 0.44

EXAMPLE 5 SodiumD-α-[DL-γ-Methylthio-α-ureidobutyramido]phenylacetamidopenicillanate.

(R = Ph; R¹ = CH₃ S (CH₂)₂ --; R³ =H; R² = NHCONH₂ ; M=Na; α¹ =DL).Prepared by Method B, from N-Carbamoyl-DL-methionine and ampicillin,isolated as the sodium salt after treatment with sodium 2-ethylhexoate.

Yield: 51%.

νmax (KBr): 3320 (broad), 1775, 1650, 1530, 1310, 1230 and 702cm⁻ ¹.

δ[(CD₃)₂ SO]: 1.48 (34.s. gem methyl); 1.58 (3H.s. gem methyl); 1.4 -2.2 (2H. m. --SCH₂ CH₂ CH<); 2.05 (3H. d. MeS-); 2.3 - 2.7 (2H. m. SCH₂--); 4.24 (1H. s. C-3 proton); 4.1 - 4.6 (1H. m.SCH₂ CH₂ CH<); 5.2 - 5.9(5H. m. β-lactams, PhCH<and -CONH₂ * ); 6.4 (1H. m. CONH*--); 7.40 (5H.m. aromatics); 8.60 and 9.00 (2 x 1H. d. CONH*--).

NH₂ OH assay: 93.8%

Biochromatography: 1 zone at R_(f) = 0.36.

EXAMPLE 6 SodiumD-α-[DL-γ-Methylthio-α-ureidobutyramido](p-hydroxyphenyl)acetamidopenicillanate.

(R = p--HQ--Ph; R¹ = CH₃ S (CH₂)₂ ; R³ = H; R² = NHCONH₂ ; M=Na; α¹=DL). Prepared by method (Bii), from N-Carbomoyl-DL-methionine andamoxycillin, isolated as the sodium salt after treatment with sodium2-ethylhexoate.

Yield: 43%.

νmax (KBr): 3350 (broad), 1770, 1650, 1510, 1235cm⁻ ¹.

δ[(CD₃)₂ SO]: 1.45 (3H.s. gem methyl); 1.57 (3H.s. gem methyl); 1.4 -2.2 (2H. m. --SCH₂ CH₂ CH<); 2.05 (3H. d. MeS-- ); 2.3 - 2.7 (2H. m.SCH₂ --); 4.24 (1H.s. C-3 proton); 4.1 - 4.6 (1H. m. SCH₂ CH₂ CH<);5.2 - 5.9 (5H.m.β-lactams, Ph CH< and CONH₂ *); 6.4 (1H. m. CONH*--);6.3 - 7.34 (4H. m. aromatics); 8.60 and 8.90 (2 × 1H.d.CONH*--).

NH₂ OH assay: 85.5%

Biochromatography: 1 zone at R_(f) = 0.29.

EXAMPLE 7 SodiumD-α-[DL-α-formamide-γ-methylthiobutyramido]phenylacetamido penicillanate

(R = Ph; R¹ = CH₃ S (CH₂)₂ ; R³ = H; R² = NHCHO; M=Na; α¹ = DL).Prepared by Method B, using N-Formyl-DL-methionine and ampicillin,isolated as the sodium salt after treatment with sodium 2-ethyl hexoate.

Yield: 63%.

νmax (KBr): 3300 (broad), 1780, 1732, 1645, 1525, 1302, 1225 and 700 cm⁻¹.

δ[(CD₃)₂ SO]: 1.46 (3H. s.gem methyl); 1.55 (3H. s.gem methyl); 1.7 -2.2 (2H. m. CH₃ SCH₂ CH₂ CH<); 2.05 (3H. d. CB₃ S--); 2.3 - 2.7 (2H. m.CH₃ S CH₂ CH₂); 4.23 (1H. s . C-3 proton); 4.70 (1H. m. --CH NHCHO);5.3 - 5.9 (3H. m. β-lactams and PhCH<); 7.37 (5H. m. aromatics); 8.08(1H. s .NHCHO); 8.31, 8.60 and 9.00 (3 × 1H. d. -CONH*--).

NH₂ OH assay: 100%

Biochromatogram: Single zone at R_(f) = 0.40.

EXAMPLE 8D-α-[DL-β-(p-Chlorophenyl)-α-ureidopropionamido]-(p-hydroxyphenyl)acetamidopenicillanicacid.

(R = p-HO-Ph; R¹ = p-Cl--PhCH₂ --; R³ = H; R² = NHCONH₂ ; M=H; α¹ = DL).Prepared by Method (Bii) from DL-β-(P-Chlorophenyl-α-ureidopropionicacid and amoxycillin.

Yield: 45%.

νmax (KBr): 3360, 1770, 1650, 1514 and 1230 cm⁻ ¹ .

δ[(CD₃)₂ SO]: 1.42 (3H. s . gem dimethyl); 1.58 (3H. s .gem dimethyl);2.86 (2H. m. --CH₂ CH<); 4.26 (1H. s . C-3 proton); 4.55 (1H. m. --CH₂CH<); 5.58 (5H. m. β-lactams, ##SPC8##

Nhconh₂ *); 6.24 (1h. m. NHCONH₂ *); 6.78 and ##SPC9##

8.53 (1H. m. CONH*); 9.04 (1H. m. CONH).

Biochromatography: R_(f) = 0.42

Analysis: C₂₆ H₂₈ N₅ O₇ SCl required: C, 52.93; H, 4.75; N, 11.87; S,5.43; Cl, 6.02. Found: C, (50.23); H, 4.79; N, 11.13; S, 5.32; Cl, 5.97.

EXAMPLE 9D-α-[DL-β-(p-Fluorophenyl)-α-ureidopropionamido]phenylacetamidopenicillanic acid

(R = Ph; R¹ = p-F-PhCH₂ ; R³ = H; R² = NHCONH₂ ; M =H; α¹ =DL) Preparedby method B from DL-β-(p-fluorophenyl)-α-ureidopropionic acid andampicillin.

Yield: 42%.

νmax (KBr): 3360, 1773, 1651, 1510, 1226 and 720 cm⁻ ¹.

δ[(CD₃)₂ SO]: 1.42 (3H.s . gem methyl); 1.57 (3H.s .gem methyl); 1.57(3H. s. gem methyl); 2.87 (2H. m. CH² CH<); 4.24 (1H.^(s) . C-3 proton);4.56 (1H. m. CH₂ CH<); 5.65 (5H. m. β-lactams, α-proton, --NHCONH₂ *),6.27 (1H. d. NHCONH₂ *); 7.24 (9H. m. Ph CH<); ##SPC10##

8.62 (1h. m. --CONH--*); 9.17 (1H. m. CONH*).

Biochromatography: 1 Spot at R_(f) = 0.42.

EXAMPLE 10D-α-[DL-β-(p-Chlorophenyl)-α-ureidopropionamido]phenylacetamidopenicillanic acid

(R = Ph; R¹ = p-Cl--PhCH₂ ; R³ = H; R² = NHCONH₂ ; M=H; α¹ = DL).Prepared by Method B, from DL-β-(p-chlorophenyl)-α-ureidopropionic acid.

Yield: 38%

ν max (KBr): 3360, 1770, 1650, 1514 and 1230 cm⁻ ¹.

δ[(CD₃)₂ SO]: 1.40 (3H.s .gem methyl); 1.56 (3H.s .gem methyl); 2.85(2H. m. --CH₂ CH<); 4.25 (1H.s . C-3 proton); 4.55 (1H. m. --CH₂ CH<);5.58 (5H. m. β-lactams, ##SPC11##

Nhconh₂); 6.25 (1h. m. NHCONH₂); 7.30 (9H.m. aromatic protons) 8.53 (1H.m. CONH); 9.00 (1H. m. CONH).

NH₂ OH Assay: 94%.

Biochromatography: 1 zone at R_(f) = 0.59.

EXAMPLE 11 SodiumD-α-[DL-β-(p-Nitrophenyl)-α-ureidopropionamido]phenylacetamidopenicillanate.

(R = Ph; R¹ = p-NO₂ --PhCH₂ ; R³ = H; R² = NHCONH₂ ; M=Na; α¹ = DL).Prepared by method B, from DL-β-(p-Nitrophenyl)-α-ureidopropionic acidand ampicillin; isolated as the sodium salt by treatment with sodium2-ethylhexoate.

Yield: 55%

ν max (KBr); 3350, 1770, 1650, 1514 and 1230 cm⁻ ¹.

δ[(CD₃)₂ SO]: 1.42 (3H. s. gem methyl); 1.60 (3H. s. gem methyl); 3.0(2H. m. -CH₂ CH<); 4.20 (1H.s. C-3 proton); 4.7 (1H. m. -CH₂ CH<); 5.3 -5.85 (5H.m. β-lactams, ##SPC12##

Nhconh₂ *); 6.2 (1h. m. NH*CONH₂); 7.18 - 8.25 (9H. m. aromatics); 8.50,9.05 (2 × 1H. m. CONH*).

NH₂ OH assay: 93.9%

Biochromatography: 1 zone at R_(f) = 0.5.

EXAMPLE 12D-α-(D-p-Phenyl-α-ureidopropionamido)-(p-hydroxyphenyl)acetamidopenicillanicacid

(R = -p--HO--Ph; R¹ = PhCH₂ ; R³ = H; R² = NHCONH₂ ; M=H; α¹ = D).Prepared by method (Biii), using D-β-phenyl-α-ureidopropionic acid andamoxycillin.

Yield: 63% M.P. 235°-238°C.

ν max (KBr): 3360 (broad), 1740, 1650, 1520 and 1230 cm⁻ ¹.

δ[(CD₃)₂ SO]: 1.47 (3H. s. gem methyl); 1.60 (3H. s. gem methyl); 2.95(2H. m. Ph.CH₂.CH<); 4.27 (1H. s. C-3 proton); 4.60 (1H. m. PhCH₂ CH<);5.30 - 5.80 (5H. m. β-lactams, ureido-NH₂ *, ##SPC13##

6.20 (1h. d. ureido-NH*--); ##SPC14##

7.25 (5h.s. benzyl aromatics), 8.50 (1H. d. --CONH--*); 9.00 (1H. d.--CONH*--).

Hydroxylamine Assay: 107.7%

Biochromatography: One zone at R_(f) = 0.35

Analysis for C₂₆ H₂₉ N₅ O₇ S; Required (%) C, 56.22; H, 5.23; N, 12.61;S, 5.77. Found (%) C, (55.33); H, 5.44; N, 11.99; S, 5.44.

EXAMPLE 13D-α-Acetamido-β-Phenylpropionamido)-Phenyl-Acetamido-Penicillanic Acid

(R = Ph: R¹ = PhCH₂ ; R³ = H; R² = NHCOCH₃ ; M = H; α¹ = DL).

Prepared by method (Bi), from DL-α-acetamido-β-phenylpropionic acid andampicillin.

Yield: 94%

ν_(max) (KBr ): 33.60 (broad) 1774, 1648, 1511, 1215 and 701cm.sup.⁻¹

δ[(CH₃)₂ SO]: 1.43 (3H.s. gem - methyl); 1.56 (3H.s. gem - methyl); 1.77(3H.s. NHCOCH₃); 2.7-3.2 (2H.m. PhCH₂ CH <) 4.24 (1H.s. C-3 proton);4.6-4.9 (1H.m PhCH₂ CH <); 5.62 (3H.m. β-lactams, PhCH <); 7.38 (10H.m.PhCH<; Ph CH₂ CH <); 8.0-9.3 (3H.m. removable in D₂ O, --CONH--)

Hydroxylamine Assay: 62.1%

Biochromatagraphy: 1 zone at R_(f) = 0.26

Analysis: C₂₇ H₃₀ N₄ O₆ S required: C, 60.22; H, 5.58; N, 10.41; S,5.95. Found: C,(57.45); H, 5.69; N, 9.98; S, 6.03.

EXAMPLE 14 D-α-[DL-α-(3-Methylureido)-β-Phenyl-Propionamido]-Phenylacetamido PenicillanicAcid

(R = Ph: R¹ = PhCH₂ ; R³ = NHCONHCH₃ ; M = H; α¹ =DL).

Prepared by method (Bi), starting from DL-α-(3methylureido)-β-phenylpropionic acid and ampicillin.

Yield: 75% ν_(max) (KBr); 1775, 1637, 1560, 1490, 1297, 1219 and702cm.sup.⁻¹. δ[(CD₃ )₂ SO]: 1.45 (3H.s. gem-methyl); 1.57 (3H.s.gem-methyl); 2.52 (3H.s. NHCONHCH₃ ); 2.90 (2H.m. PhCh₂ CH<); 4.27(1H.s. C-3 proton); 4.63 (1H.m. PhCH₂ CH<); 5.64 (3H.m. β-lactams, PhCH₂<); 7.32 (10H.m. PhCH₂ CH<; Ph CH<); 8.63 (1H.m. removable in D₂O,--CONH --); 9.18 (1H.m. removable in D₂ O --CONH --); ca 6.25 (broadsignal due to --NH CONH--).

Biochromatography: 1 zone at R_(f) = 0.47

Analysis: C₂₇ H₃₁ N₅ O₆ S required: C, 58.48, H, 5.60; N, 12.64; S, 5.78Found: C, (56.78); H, 5.59; N, 12.73; S, 5.05

EXAMPLE 15D-α-[DL-β-(-p-Fluorophenyl)-α-Ureido-Propionamido]-(-p-Hydroxyphenyl)-Acetamido-PencillanicAcid

(R = -p-HO--Ph; R¹ = --p-F--PhCH₂ ; R³ = H; R² = NHCONH₂ ; M = H, α¹ =DL)

Prepared by method (Bii), from DL-β-(-p-fluorophenyl)-α-ureidopropionicacid and amoxycillin.

Yield: 48%

ν_(max) (KBr): 3360, 1764, 1650, 1510, 1224, and 838cm.sup.⁻¹

δ[(CD₃)₂ SO]: 1.43 (3H.s. gem-methyl); 1.54 (3H.s. gem-methyl); 2.88(2H.m. -CH₂ CH<); 4.14 (1H.s. C-3 proton); 4.2-4.8 (1H.M. CH₂ CH<);5.3-7.5 (extremely strong signals containing β-lactams, ##SPC15##

--NHCONH₂ *, ##SPC16##

δ = 8.3-9.2 (2h.m. --CONH-- *)

Hydroxylamine Assay: 84.7%

Biochromatography: 1 zone at R_(f) = 0.41

EXAMPLE 16 D-α-[D-β-Phenyl-α-(n-Valeramido)-Propionamido]Phenylacetamido Penicillanic Acid

(R = Ph; R¹ = PhCH₂ ; R³ = H; R² = NHCO (CH₂)₃ CH₃ ; M = H; α¹ = D).

Prepared by method (Bi), from N -valeroyl-D-β-phenylalanine

Yield: 19% ν_(max) (KBr): 3290 (br), 1772, 1635, 1525, 1300, 1224, 733,702^(cm) ⁻¹. δ[(CD₃)SO]; 0.82 (3H.m. (CH₂)₃ CH₃), 1.0-1.7 (4H.m. CH₂ CH₂CH₂ CH₃) 1.45 (3H.s. gem-methyl), 1.58 (3H.s. gem-methyl), 2.09 (2H.m.CH₂ (CH₂)₂ CH₃), 3.00 (2H.m. PhCH₂), 4.28 (1H.s. C-3 proton), 4.78(1H.m. PhCH₂ CH<) 5.4-5.9 (3H.m. β-lactams and PhCH<), 7.2-7.5 (10H.m.aromatic protons), 8.05 (1H.d. --CONH--), 8.47 (1H.m. CONH), 9.13 (1H.m.--CONH--).

NH₂ OH Assay: 90%

Biochromatography: Single zone R_(f) 0.70

Analysis: Found: C, 60.96, H, 6.05; N, 9.46; S, 5.65% C₃₀ H₃₆ N₄ O₆ Srequires C, 62.10; H, 6.21; N, 9.66; S, 5.52

EXAMPLE 17 D-α-[D-β-Phenyl-α-Pivaloylaminopropionamido]-PhenylacetamidoPenicillanic Acid

(R = Ph; R¹ = PhCH₂ ; R³ = H; R² = --NHCOC(CH₃)₃ ; M = H; α¹ = D).

Prepared by method (Bi), from α-t-Butyramido-D-β-phenyl propionic acid.

Yield: 6%

ν_(max) (KBr): 3350(br), 1772, 1639, 1517, 1300, 1212, 702cm⁻ ¹.

δ[(CD₃)₂ SO] : 0.97 (9H.s. (CH₃)₃), 1.39 (3H.s. gem-methyl), 1.52 (3H.s.gem-methyl), 2.97 (2H.m. PhCH₂), 4.20 (1H.s. C-3 proton), 5.35-5.85(4H.m. β-lactams and PhCH<), 4.65 (1H.m. PhCH₂ CH<), 7.25 (5H.s. phenylprotons), 7.39 (5H.m. phenyl protons), 7.2-7.6 (1H.m. --NHCO--*), 8.50(1H.d. --NHCO--*), 9.27 (1H.d. --NH--CO--).

NH₂ OH Assay: 102%

Biochromatography: 0.71

EXAMPLE 18 D-α-[D,L-α-Benzamido-β-Phenylpropionamido]-PhenylacetamidoPenicillanic Acid

(R = Ph; R¹ = PhCH₂ ; R³ = H; R² = NHCOPh; M = H; α¹ = D,L). Prepared bymethod (Bi), from α-Benzamido-D,L-β-phenylpropionic acid.

Yield: 22%

ν_(max) (KBr): 3300, 1775, 1635, 1522, 1302, 122, 702cm⁻ ¹

δ[(CD₃)₂ SO] : 1.42 (3H.s. gem-methyl), 1.57 (3H.s. gem-methyl) 3.11(2H.m. PhCH₂), 4.24 (1H.s. C-3 proton), 5.00 (1H.m. PhCH₂ CH<), 5.4-5.9(3H.m. β-lactams and PhCH<), 7.2-8.0 (10H.m. aromatic protons), 8.65,8.80 and 9.20 (3 × 1H.d. --NHCO--*).

NH₂ OH Assay: 88%

Biochromatography: Single zone 0.70

EXAMPLE 19D-α-[D,L-γ-Phenyl-α-Ureidobutyramido]-p-HydroxyphenylacetamidoPenicillanic Acid

(R = p HO--Ph; R¹ = PhCH₂ CH₂ --; R³ =H; R² = --NHCONH₂ M = H; α¹ =D,L). Prepared by method (Biii), from α-ureido-D,L-α-phenylbutyric acid.

Yield: 35%

ν_(max) (KBr): 3315(br), 1770, 1650, 1510, 1454, 1227, 842, 703cm.sup.⁻¹

δ[(CD₃)₂ SO]: 1.42 (3H.s. gem-methyl), 1.52 (3H.s. gem-methyl), 2.00(2H.m. PHCH₂ CH₂ ), 2.52 (2H.m. PhCH₂ CH₂), 4.1-4.4 (1H.m. PhCH₂ CH₂CH<), 4.25 (1H.s. C-3 proton), 5.3-5.8 (3H.m. β-lactams and PhCH<), 7.25(9H.m. aromatic protons), 6.34, 6.73 (2 × 1H.d. NHCO*), 8.32-9.10 (3H.m.--CONH--* and --CONH₂ *)

NH₂ OH Assay: 80%

Biochromatography: R_(f) 0.5

EXAMPLE 20 D-α-[D,L-α- Formamido-β-Phenylpropionamido]-PhenylacetamidoPenicillanic Acid

(R = Ph; R¹ = PhCH₂ ; R³ = H; R² = --NHCHO; M = H; α¹ = D,L). Preparedby method (Bi) from N-Formyl-D,L-phenylalanine.

Yield: 55%

ν_(max) (KBr): 3242(br), 1771,,1638, 1522, 1379, 1300, 1226, 731, 701cm⁻¹.

δ[(CD₃)₂ SO]: 1.46 (3H.s. gem-methyl), 1.59 (3H.s. gem-methyl); 2.88(2H.m. PhCH₂ CH<), 4.21 (1H.s. C-3 proton), 4.83 (1H.m. PhCH₂ CH<),5.4-5.9 (3H.m. β-lactams and PhCH<), 7.2-7.6 (10H.m. aromatic protons),7.97 (1H.s. CHO), 2.27, 2.70 and 9.11 (3 × 1H.d. --NHCO*--).

NH₂ OH Assay: 79%

Biochromatography: Single zone R_(f) 0.52

EXAMPLE 21 D-α-[D-β-Phenyl-α-Propionamido Propionamido]-PhenylacetamidoPenicillanic Acid

(R = Ph; R¹ = PhCH₂ ; R³ = H; R² = NHCOCH₂ CH₃ ; M = H; α¹ = D).Prepared by method (Bi) from α-Propionamido-D-β-phenylpropionic acid.

Yield: 13%

ν_(max) (KBr): 3229(br), 1770, 1637, 1524, 1226, 702cm.sup.⁻¹.

δ[(CD₃)₂ SO]: 0.89 (3H.t. COCH₂ CH₃), 1.42 (3H.s. gem-methyl), 1.60(3H.s. gem-methyl), 1.98 (2H.m. --NHCH₂ CH₃), 2.90 (2H.m. PhCH₂ CH<),4.21 (1H.s. C-3 proton), 4.70 (1.H.m. PhCH.sub. 2 CH<), 5.4-5.9 (3H.m.β-lactams and PhCH <), 7.2-7.6 (10H.m. aromatic protons), 8.01, 8.42 and9.10 (3 × 1H.d. --CONH*--).

NH₂ OH Assay: 75%

Biochromatography: Single zone R_(f) 0.58

EXAMPLE 22 D-α-[D-α-Isobutyramido-β-Phenylropionamido]-PhenylacetamidoPenicillanic Acid

(R = Ph; R¹ = PhCH₂ --; R³ = H; R² = NHCOCH(CH₃)₂ ; M = H; α¹ = D.Prepared by method (Bi), from α-isobutyramido-D-β-phenyl propionic acid

Yield: 18%

ν_(max) (KBr): 3300 (br,) 1771, 1638, 1526, 1300, 1222, 702cm⁻ ¹.

δ[(CD)₂ SO]: 0.85 (6H.t. CH(CH₃)₂, 1.41 (3H.s. gem-methyl), 1.56 (3H.s.gem-methyl), 2.97 (2H.m. PhCH₂ CH<), 4.25 (1H.s. C-3 proton), 4.70(2H.m. CH (CH₃)₂ and PhCH₂ CH<), 5.4-5.9 (3H.m. β-lactams and PhCH<),7.1-7.6 (10H.m. aromatic protons), 7.97, 8.47 and 9.13 (3 × 1H.d. --NHCO--*).

NH₂ OH Assay: 103%

Biochromatography: Single zone R_(f) 0.66

EXAMPLE 23 D-α-[D-α-Methylthio-γ-Ureidobutyramido]-PhenylacetamidoPenicillanic Acid

(R = Ph; R¹ = CH₃ S(CH₂)₂ --; R³ = H; R² = --NHCONH₂ ; M = H; α¹ = D).Prepared by method (Fi), from N-Carbamoyl-D-methionine.

Yield: 43%

ν_(max) (KBr): 3320(br), 1775, 1650, 1530, 1310, 1230, and 702cm⁻ ¹

δ[(CD₃)₂ SO]: 1.48 (3H.s. gem-methyl), 1.61 (3H.s. gem-methyl), 1.4-2.2(2H.m. -SCH₂ CH₂ CH<), 2.1 (3H.s.CH₃ S--), 2.3-2.7 (2H.m. --SCH₂ CH₂ CH)5.2-5.9 (5H.m.β-lactams, PhCh and --CONH₂ *) 6.4 (1H.m. --CONH --*)

NH₂ OH Assay: 98%

Biochromatography: Single zone R_(f) 0.34

Analysis: Found: C, 49.33; H, 5.64; N, 12.94; S, 11.59%. C₂₂ H₂₉ N₅ O₆S₂ requires C, 50.48; H, 5.54; N, 13.38; S, 12.24%.

EXAMPLE 24 D-α-[D,L-α-Methyl-α-Ureidovaleramido]-PhenylacetamidoPenicillanic Acid

(R = Ph; R¹ = (CH₃)₂ CHCH₂ --; R³ = H; R² = --NHCONH₂ ; M = H; α¹ =D,L)

Prepared by method B from α-methyl-D,L-α-ureido valeric acid.

Yield: 36%

ν_(max) (KBr): 3325(br), 1775, 1723, 1650, 1530, 1310, 1222, 702cm⁻ ¹δ[(CD₃)₂ SO]: 0.84 and 0.95 (2 × 3H.s. CH(CH₃)₂), 1.45 (3H.s.gem-methyl), 1.61 (3H.s. gem-methyl), 0.78-2.0 (3H.m. CH₂ CH(CH₃)₂),4.25 (1H.s. C-3 proton), 4.0- 4.6 (1H.m. >CHNHCONH₂), 5.3- 5.9 (5H.m.β-lactams, PhCH< and CONH₂ *), 6.2 (1H.d. --CONH--), 7.2- 7.6 (5H.m.aromatic protons), 8.48 and 9.04 (2 × 1H.d. --CO--NH--*)

NH₂ OH Assay: 79%

Biochromatography: Single zone R_(f) 0.44

EXAMPLE 25 D-α[ D,L-α-Formamido-β-(p-hydroxyphenyl)propionamido]phenylacetamido penicillanic acid

(R=Ph; R'=pHO--PhCH₂ --; R³ = H; R² =NHCHO; M=H; α'=D,L). Prepared bymethod B from N-Formyl-D,L-Tyrosine.

Yield: 23%

ν_(max) (KBr): 3290 (br.), 1773, 1735, 1650, 1518, 1378, 1230 and 701cm⁻ ¹

δ[ (CD₃)₂ SO] 1.45 (3H.s gem. methyl), 1.62 (3H.s.gem.methyl), 2.7-3.2(2H.m. PhCH₂ --), 4.24 (1H.s C3 proton), 4.8 (1H.m. >CHNHCHO), 5.35-5.96(3H.m βLactams and ph CH<), 6.7 and 7.0 (2 × 2H.m. pHO--C₆ H₄ --), 7.36(5H.m. Aromatic protons), 7.92 (1H.s. --NHCHO), 8.2 (1H.d --CONH--*) 8.7and 9.12 (2 × 1H.m.--CONH--*).

NH₂ OH Assay: 105%

Biochromatogram: Single zone R_(f) 0.39.

EXAMPLE 26 D-α-[D-α-Formamido-γ-methylthiobutyramido]-phenylacetamidopenicillanic acid.

(R=Ph; R'=CH₃ S (CH₂)₂ --;R³ =H; R² =--NHCHO; M=H; α'=D). Prepared bymethod (Fii) from N - Formyl-D-methionine.

Yield: 5.8%

ν_(max) (KBr): 3300 (br), 1780, 1732, 1645, 1525, 1302, 1225, 700 cm⁻ ¹.

δ [ (CD₃)₂ SO]: 1.46 (3H.s gem. methyl), 1.55 (3H.S. gem methyl), 1.7-22 (2H.m. CH₃ SCH₂ CH₂ CH<), 2.0 (s. 15- 17% of L-CH₃ SCH₂ CH₂ C*H<) 2.1(S.83- 5% of D-CH₃ SCH₂ CH₂ C*H<), 2.3-2.7 (2H.m. CH₃ SCH₂ CH₂ CH₂),4.23 (1H.S. C₃ proton), 4.70 (1H.m. >CHNHCHO), 5.3-5.9 (3H.m. β-lactams,and PhCH<), 7.37 (5H.m. aromatic protons), 8.08 (1H.s-NH-CHO), 8.31,8.59 and 9.02 (3 × 1H.d.--CONH-- removed by D₂ O).

NH₂ OH Assay: 102%

Biochromatogram: Single zone Rf 0.41

EXAMPLE 27 D - α-[D-γ-Methyl-α-ureidovaleramido]-phenylacetamidopenicillanic acid.

(R=Ph; R'= (CH₃)₂ CHCH₂ ; R.sup. 3 =H; R² =NHCONH₂ ; M=H; α'=D) Preparedby method (Fi) from γ-methyl-D-α-ureido valeric acid.

Yield: 28% M.p: 168-170°C (Decomp.)

ν_(max) (KBr): 3315 (br.), 1775, 1730, 1650, 1530, 1310, 1220, 700 cm⁻¹.

δ[(CD₃)₂ SO]: 0.85 and 0.92 (6H.d. CH(CH₃)₂), 1.50 (3H.S gem. methyl)1.66 (3H.S gem.methyl), 0.8-2.0 (3H.m. CH₂ CH (CH₃)₂), 4.28 (1H.s. C3proton), 4.1-4.5 (1H.m. CH NHCONH₂), 5.3-5.9 (5H.m. β-lactams PhCH< andCONH*₂), 6.22 (1H.d.--CONH*--), 7.36 (5H.m. aromatic protons), 9.07 and9.40 (2 × 1H.d. -- CO NH*-- )

NH₂ OH Assay: 95%

Biochromatogram: Single zone Rf0.45

Analysis: Found: C,54.71; H, 6.15; N,13.78; S,6.49%. C₂₃ H₃₁ N₅ O₆ Srequires C, 54.65, H, 6.14; N, 13.86; S,6.34%.

EXAMPLE 28 D-α-[L-γ-Methylthio-α-ureidobutyramido] phenylacetamidopenicillanic acid

(R=Ph; R'= (CH₃) S (CH₂)₂ --; R² =H; R² = NHCONH₂ ; M=H, α'=L). Preparedby method (Fi) from N-Carbamoyl-L-methionine.

Yield: 19% M.p.: 163°-6°C (Decomp.)

ν_(max) (KBr): 3350 (br), 1775, 1650, 1522, 1305, 1220, 702 cm⁻ ¹.

δ[(CD₃)₂ SO]: 1.46 (3H.s gem.methyl), 1.60 (3H.s.gem.methyl) 1.5-2.2(2H.m. CH₃ CH₂ CH₂ CH<), 2.00 (3H.s.CH₃ S) 2.2-2.6 (2H.m. CH₃ SCH₂ CH₂CH), 4.27 (1H.S. C3 proton), 4.38 (1H.m. CH₃ CH₂ CH₂ CH<), 5.60 (5H.m βlactams, PhCH< --CONH*₂), 7.39 (5H.m. aromatic protons), 6.34 (1H.m.--CHCONH*₂), 9.12 and 9.48 (2 × 1H.d --CONH*--)

NH₂ OH Assay: 100%

Biochromatogram: Single zone Rf0.38

Analysis: Found: C,49.64; H, 5.98; N,13.08; S,11.72%. C₂₂ H₂₉ N₅ O₆ S₂requires C 50.48; H,5.54; N,13.38; S,12.24%

EXAMPLE 29 Triethylammonium D-α-[D-β-(3-Indolyl)-α-ureido-propionamido]phenylacetamido penicillanate

(R=Ph; ##SPC17##

R³ =h; r² =nhconh₂ ; m=hn (ch₂ ch₃)₃ ; α'=d. prepared by method (Fiii)from N - Carbamoyl-D-Tryptophan

Yield: 52% M.p.: 200°-3°C (Decomp).

ν_(max) (KBr): 3350 (br), 1767, 1660, 1640, 1610, 1530, 1458, 1392, 749cm⁻ ¹.

δ[(CH₃)₂ SO]: 1.11 (3H.t.OCH₂ CH₃), 1.42 (3H.s. gem. methyl), 1.53 (3H.s.gem methyl), 2.7-3.2 (4H.m.-CH₂ CH< and OCH₂ CH₃), 4.00 (1H.s. C₃proton), 4.59 (1H.m. CH₂ CH<) 5.3-5.9 (5H.m. β lactams, RhCh< andCONH*₂), 6.9-7.7 (10H.m. aromatic protons), 6.29, 8.53 and 8.97 (3 ×1H.d. --NHCO*), 10.84 (1H.s. indolyl NH* ).

NH₂ OH Assay: 94%

Biochromatogram: Single zone Rf0.30

Analysis: Found: C,59.38; H,6.65; N, 14.35; S,4.60% C₃₄ H₄₅ N₇ O₆ Srequires C,60.07; H,6.67; N,14.42, S,4.72%

EXAMPLE 30 TriethylammoniumD-α-[D,L-α-formamido-β-(3-indolyl)-propionamido]phenylacetamidopenicillinate

(R= Ph: ##SPC18##

R³ =h; r² =--nhcho;m=hn (ch₂ ch₃)₃ ; α'=d,l. prepared by method (Fi)from N - Formyl-D,L-tryptophan, the product crystallising on dilutionwith ether.

Yield: 50%

ν_(max) (KBr): 3310 (br), 1772, 1770, 1665, 1530, 1458, 1388, 1218, 748cm⁻ ¹.

δ[(CD₃) SO]: 1.17 (3H.t.OCH₂ CH₃), 1.44 (3H.s. gem.methyl), 1.58 (3H.s.gem methyl), 2.8-3.3 (4H.m. OCH₂ CH₃ and Ch₂ CH<), 4.12 (1H.s. C₃proton), 4.90 (1H.m. CH₂ CH<), 5.3 - 5.9 (3H.m. β-lactams and Ph CH<),7.0-7.8 (11H.m. 10 aromatic protons and NHCONH₂ ) 8.00 (1H.S. CHO),8.0-9.0 (4H.m. 2 ×-NHCO* and --CONH₂ *), 10.80 (1H.s indolyl NH*)

NH₂ OH Assay: 183%

Biochromatogram: Single zone Rf0.36.

EXAMPLE 31 Triethylammonium -α-[D-γ-carbamoyl-α-Ureido butyrylamido]phenylacetamido penicillanate

(R=Ph; R'=H₂ NCO (CH₂)₂ --; R³ =H; R² =--NHCONH₂ ; M=HN (CH₂ CH₃)₃ ;α'=D. Prepared by method (Fi) from N-carbamoyl-D-glutamine, the productcrystallising on dilution with ether.

Yield: 77%

ν_(max) (KBr): 3400 (br.), 1773, 1698, 1660, 1603, 1532, 1458, 1397,1314, 1220, 703 cm⁻ ¹.

δ[(CD₃)₂ SO]: 1.16 (3H.t.OCH₂ CH₃), 1.43 (3H.s. gem. methyl), 1.54(3H.s. gem methyl), 1.5-2.2 (4H.m. -- CH₂ CH₂ --), 4.1(1H.s.C3proton),4.88 (1H.m. CH Ch₂ CH₂), 5.3-5.8 (3H.m β-lactams and PhCH<), 6.3(1H.m.--NHCO*--), 7.3-7.7 (6H.m. aromatic protons and --NHCO*--),8.2-9.0 (5H.m. 2 × --CONH*₂ ) and --CONH*

NH₂ OH Assay: 100%

Biochromatogram: Single zone Rf0.34

EXAMPLE 32D-α-[D,L-β-Phenyl-α-ureidopropionamido]-1,4-cyclohexadienylacetamidopenicillanicacid. ##SPC19##

R³ =h; r² =--nhconh₂ : m=h; α¹ =d,l). prepared by method (Bv) fromD,L-β-phenyl-α-ureidopropionic acid.

Yield: 65%

ν_(max) (KBr): 3340, 1762, 1720, 1650, 1530, 1230 and 703cm⁻ ¹

δ [(CD₃)₂ SO]: 1.5(6H.d. gem-dimethyls); 2.6(4H.s. cyclohexadienemethylenes); 2.6-3.2(2H.m. PhCH₂ CH<); ); 4.3(1H.s.C-3proton);4.3-4.8(1H.m. PhCH₂ CH<); 4.9-5.9 (5H.m.β-lactams, ##SPC20##

--NHCONH₂ *); 5.67(3H.s. cyclohexadiene methines); 6.3-6.7(1H.m.-NHCONH₂ *);7.25(5H.s.PhCH₂ CH<); 8.0-8.3(1H.m.--CONH--*);8.6-9.0(1H.m.--CONH--*);

Hydroxylamine Assay: 59%.

Biochromatography: Rf 0.56.

EXAMPLE 33 SodiumD-β-[D-β-Phenyl-β-ureidopropionamido]-1,4-cyclohexadienylacetamidopenicillanate ##SPC21##

R¹ =phCH₂ ; R³ =H; R² =--NHCONH₂ ; M=Na; α¹ =D) Prepared by method (Bvi)from D-β-phenyl-α-ureidopropionic acid and epicillin.

Yield: 45%.

ν_(max) (KBr): 3350, 1760, 1630, 1530, 1230 and 703cm⁻ ¹

δ[(CD₃)₂ SO]: 1.53(6H.d.gem-dimethyls); 2.65(4H.s.cyclohexadienylmethylenes); 2.8-3.0(2H.m.PhCH₂ --);4.0(1H.s.C-3 proton);4.2-4.7(1H.m.)CH₂ CH--);5.0-5.6(5H.m.β-lactam protons HNCHCO andHNCONH₂):5.7(3H.s. cyclohexadienyl vinylic protons); 6.4-6.7(1H.m.-NHCONH₂); 7.25(5H.s.Ph); 8.0-9.0(2H.m.NH)

Hydroxylamine Assay: 76%.

Biochromatography: 1 zoneat Rf 0.47

EXAMPLE 34D-α-[DL-β-Benzyloxy-α-ureidopropionamido]-phenylacetamidopenicillanicacid.

(R=Ph; R¹ =PhCH₂ OCH₂ ; R³ =H; R² =NHCONH₂ ; M=H; α¹ =DL). Prepared bymethod (Bi) from β-benzyloxy-α-ureido-DL-propionic acid and ampicillin.

Yield: 56%.

ν_(max) (KBr): 3350, 1770, 1650, 1520, 1220 and 700cm⁻ ¹

δ[(CD₃)₂ SO]: 1.5(6H.d.gem dimethyls); 3.65(2H.m. --OCH₂ CHC<)4.25(1H.s.C-3 proton); 4.5(3H.m. --OCH₂ CH<and PhCH₂ O--); 5.4-7.0(6H.m.β-lactam protons, HN--CHCO and --NHCONH₂); 7.2-7.5(10H.m. 2xphenylaromatics); 8.5-9.2(2H.m. amide NH's)

Analysis: C₂₂ H₃₁ N₅ O₇ S.H₂ O required: C,55.3; H,5.63: N, 11.9. Found:C,56.03;H,5,72: N,11.46.

Hydroxylamine Assay: 95%.

Biochromatography: 1 zone at Rf 0.42.

EXAMPLE 35D-α-[DL-β-Benzyloxy-α-ureidopropionamido]-p-hydroxyphenylacetamidopenicillanic acid.

(R= p-HO--Ph; R¹ =PhCH₂ OCH₂ ; R³ =H; R² =--NHCONH₂ ; M=H; α¹ =DL).Prepared by method (Biii) from DL-β-benzyloxy-α-ureidopropionic acid andamoxycillin.

Yield: 59%.

ν_(max) (KBr): 3350, 1775, 1725, 1650, 1515, 1230, and 703 cm⁻ ¹

δ[(CD₃)₂ SO]: 1.5(6H.d. gem dimethyls); 3.5-3.9(2H.m. --OCH₂ CH<); 4.23(1H.s. C-3 proton); 4.3-4.7(3H.m. PhCH₂ O and OCH₂ CH<); 5.2-5.9(5H.m.β-lactams, HNCHCO and CONH₂) 6.2-6.5 (1H.m. -NHCONH₂); 6.5-7.5(9H.m.aromatic protons) 8.3-9.0 (2H.m. amide NH's)

Hydroxylamine Assay: 84%.

Biochromatography: 1 zone at Rf 0.3

EXAMPLE 36 D-α-[D-β-Phenyl-α -ureidopropionamido]-3-thienyl acetamidopenicillanic acid ##SPC22##

R¹ =phch₂ -; r³ =h; r² =-nhconh₂ ; m=h, α¹ =d). prepared by method (Biv)from D-β-phenyl -α-ureidopropionic acid.

Yield: 59%. M.p. 175°-7°C (decomp.)

ν_(max) (KBr): 3360, 1750, 1650, 1525 and 704 cm⁻ ¹

δ[(CD₃)₂ SO]: 1.53(6H.m. gem dimethyls); 2.92(2H.m. PhCH₂ CH<)4.28(1H.s. C-3proton); 4.61(1H.m. PhCH₂ CH<); 5.31-6.05 (5H.m.β-lactams, CONH₂ * and ThCH <); 6.26(1H.d. CONH--*); 7.37(8H.m. Ph andTh); 8.58 and 9.07 (2×1H.d. --CONH--*)

Hydroxylamine Assay: 100%.

Biochromatogram : 1 zone at Rf 0.40

EXAMPLE 37 D-β-[D-α-Ureidopropionamido]-phenylacetamido penicillanicacid.

(R=Ph; R¹ =CH₃ ; R³ =H; R² =NHCONH₂ ; M=H; α¹ =D). Prepared by method(Bi) from D-α-ureidopropionic acid.

Yield: 28%. M.p. 176°-8°C (decomp.)

ν_(max) (KBr): 3350 (br), 1773, 1720, 1635, 1530, 1234, and 700 cm⁻ ¹

δ [(CD₃)₂ SO]: 1.21 (3H.d. CH₃ CHNH--); 1.43 (3H.s. gem dimethyls); 1.57(3H.s. gem dimethyls); 4.26 (1H.s. C-3 proton); 4.33(1H.m. CH₃ CHNH--);5.35-5.91(5H.m. β-lactams, PhCH< and CONH₂ *); 5.91-6.54 (1H.m.--NHCONH₂); 7.38(5H.m. aromatic protons); 8.47 and 9.05 (2x1H.d.--CONH--*)

Hydroxylamine Assay: 106%.

Biochromatogram: 1 zone at Rf 0.21

EXAMPLE 38 D-α-[D,L-β-PHENYL-α-UREIDO-PROPIONAMIDO]-PHENYLACETAMIDOPENICILLANIC ACID

(R = Ph ; R¹ = PhCH₂ R³ = H ; M = H ; α¹ = D,L).

Prepared by method B from D,L-β-Phenyl-α-ureido propionic acid.

Yield: 37%

ν_(max) (KBr): 3350 (br.), 1775, 1650, 1525, 1225, 702cm⁻ ¹.

δ[(CD₃)₂ SO]: 1.45 (3H.s. gem methyl), 1.57 (3H.s. gem methyl), 2.88(2H.m. PhCH₂ --), 4.26 (1H.s. C3proton), 4.60 (1H.m. PhCH₂ CH<),5.33-5.94(5H.m. β-lactams, PhCH,--CONH₂ *), 6.24 (1H.d.--CONH*--), 7.27(10H.m. aromatic protons), 8.54 (1H.d.--CONH*--), 9.11 (1H.d.--CONH*--)

NH₂ OH Assay: 87%

Biochromatogram: Single zone R_(f) 0.42.

EXAMPLE 39 D-α-[-α-Methyl-α-Ureidopropionamido]-PhenylacetamidoPenicillanic Acid

(R = Ph ; R¹ = R³ = CH₃ --; R² = NH₂ CONH-- ; M = H). Prepared by methodB from α-Ureido-isobutyric acid

Yield: 6%

ν_(max) (KBr): 3400 (br.), 1785, 1715, 1650, 1535, 1225 and 700Cm⁻ ¹

δ[(CD₃)₂ SO]: 1.38 (6H.s. (CH₃)₂ C<), 1.46 (3H.s. gem methyl) 1.57(3H.s. gem methyl), 4.27 (1H.s. C3 proton), 5.37-5.88 (5H.m. β-lactams,CONH₂ *, Ph CH<), 6.37 (1H.s. --NHCO*--), 7.40 (5H.m. aromatic protons),8.13 and 9.08 (2 × 1H.d.--CONH*--)

NH₂ OH Assay: --

Biochromatogram: Single zone R_(f) 0.27

Analysis: Found; C,52.60; H,5.89; N,14.34; S,6.88% C₂₁ H₂₇ N₅ O₆ Srequired C,52.82; H,5.70; N,14.67; S,6.71%

EXAMPLE 40D-α-[D,L-α-Methyl-β-Phenyl-α-Ureidopropionamido]-PhenylacetamidoPenicillanic Acid

(R = Ph; R¹ = PhCH₂ --; R³ = CH₃, R² = NHCONH₂, α' = D,L)

Prepared by method B from D,L-α-Methyl-β-phenyl-α-ureidopropionic acid.

Yield: 43%

ν_(max) (KBr): 3370 (br.), 1775, 1720, 1655, 1525, 1220 and 704 cm⁻ ¹.

δ[(CD₃)₂ SO]: ##EQU5## 1.43 (3H.s. gem. methyl), 1.57 (3H.s. gemmethyl), 2.80-3.70(2H.m.PhCH₂₋₋), 4.30 (1H.m. C3 proton), 5.38-6.12(5H.m. β-lactams, PhCH and CONH₂ * ), 6.23 (1H.s.--CONH--), 7.39 (10H.m.aromatic protons), 8.34(1H.d.--CONH--*), 9.28(1H.d.-CONH--*) * removedby D₂ O

Biochromatogram: Single zone R_(f) 0.54.

Analysis: Found; C56.49; H,5.66; N,12.16%. C₂₇ H₃₁ N₅ O₆ S requiresC,58.58; H,5.64; N,12.65%

EXAMPLE 41D-α-[D,L-α-Acetamido-βPhenylpropionamido]-p-HydroxyphenylacetamidoPENICILLANIC ACID

(R = p HO-Ph; R¹ = PhCH₂ ; R³ = H ; R² = --NHCOCH₃ ; M = H; α¹ = D,L).

Prepared by method (Bii) from N-Acetyl-D,L-β-phenylalanine.

YIELD: 40%

ν_(max) (Nujol): 3250 (br.), 1760, 1630, 1515, 1380, 1220, 710cm⁻ ¹.

δ[(CD₃)₂ SO]: 1.43 (3H.s. gem methyl), 1.57 (3H.s. gem methyl) 1.75(3H.s. --NHCOCH₃), 3.0 (2H.m. PhCH₂), 4.24 (1H.s. C3 proton), 6.72(1H.m. PhCH₂ CH<), 5.4-5.9 (3H.m. β-lactams and PhCH<), 6.58-7.50 (9H.m.aromatic protons), 8.10, 8.47 and 8.95 (3 × 1H.m. --CONH--*).

NH₂ OH Assay: 105%

Biochromatogram; Single zone R_(f) 0.51

EXAMPLE 42D-α-[L-β-(p-Methoxyphenyl)-α-Ureidopropionamide]PhenylacetamidoPenicillanic Acid

(R = Ph ; R₁ = pMeO--PhCH₂ -- ; R³ = H; R² = --NHCONH₂ ; M = H; α¹ = L).

Prepared by method B from α Ureido-L-β-(p Methoxyphenyl) propionic acid.

Yield: 19%

ν_(max) (KBr): 3300 (br.), 1775, 1640, 1515, 1250 and 700cm⁻ ¹.

δ[(CD₃)₂ SO]: 1.45 (3H.s. gem methyl), 1.58 (3H.s. gem methyl), 2.83(2H.m. PhCH₂), 3.72(3H.s. CH₃ O), 4.28 (1H.s. C3 proton), 4.61 (1H.m.PhCh₂ CH), 5.35-5.93 (5H.m. β-lactams, PhCH, --NHCONH₂ *), 6.27 (1H.d.--NHCO--), 6.67-7.50 (9H.m. aromatic protons), 8.70 and 9.32 (2 × 1H.d.--NHCO--*)

NH₂ OH Assay: 81%

Biochromatogram: Single zone R_(f) 0.43.

EXAMPLE 43 D-α-[D-α-Guanidino-β-Phenylpropionamido]-PhenylacetamidoPenicillanic Acid Hydrochloride

(R = Ph ; R¹ =PhCH₂ --; ##EQU6## M = H α¹ = D).

Prepared by method E from D-α-Guanidino-β-phenyl propionic acid.

Yield: 21%

ν_(max) (KBr): 3330 (br.), 1768, 1663, 1602, 1525, 1458 1394, 1320,703cm⁻ ¹.

δ[(CD₃)₂ SO]: 1.44 (3H.s. gem methyl), 1.57 (3H.s. gem methyl),2.96-3.23 (2H.m. PhCH₂ --), 4.17(1H.m. C3 proton), 4.61 (1H.m. PhCH₂CH<), 5.30-5.94 (3H.m. β-lactams and PhCH<), 7.11-7.71 (14H.m. aromaticprotons and ##EQU7## 8.14-8.47 8.82 and 9.12 (2 × 1H.m. --CONH--*)

NH₂ OH Assay: 90%

Biochromatogram: Single zone R_(f) 0.52

EXAMPLE 44 D-α-[D,L-β-Methoxy-α-Ureidopropionamido]-Phenyl-AcetamidoPenicillanic Acid

(R = Ph ; R¹ = CH₃ OCH₂ ; R³ = H ; R² = --NHCONH₂ ; M = H ; α = D,L).

Prepared by method B from D-α-Ureido-β-methoxy propionic acid.

Yield; 23%

ν_(max) (KBr): 3350 (br.), 1775, 1650, 1520, 1310, 1225, 1115 and 700cm⁻¹.

δ[(CD₃)₂ SO]: 1.45 (3H.s. gem methyl), 1.58 (3H.s. gem methyl), 3.27(3H.d. CH₃ OCH₂ --), 3.57 (2H.m. CH₃ OCH₂ --), 4.18-4.75 (1H.m. --CH₂CH<), 4.27 (1H.s. C3proton), 5.23-5.98(5H.m. β-lactams, --NHCONH₂ *,PhCH<), ), 6.37 (1H.d.--NHCO--*), 7.38 (5H.m. aromatic protons) 8.42(1H.d. --CONH--*), 9.14 (1H.m.--CONH--*).

NH₂ OH Assay: 98%

Biochromatogram: Single zone R_(f) = 0.24

EXAMPLE 45 PHTHALID-3-yl D-α-[D-α-GUANIDINO -β-PHENYLPROPIONAMIDO]PHENYLACETAMIDO PENICILLINATE, HYDROCHLORIDE

(R = Ph; R¹ = PhCH₂ - ; R³ = H ; ##EQU8## ##SPC23## α¹ = D).

Prepared by method A from D-α-Guanidino-β-phenyl propionic acid.

Yield: 23%

νmax (KBr): 3340 (br.), 1785, 1660, 1510, 1285, 980 755 and 705cm⁻ ¹.

δ[(CD₃)₂ SO]: 1.53 (6H.m. gemdimethyls), 3.03 (2.H.m. PhCH₂ CH), 4.56(1H.s. C3 proton), 4.37-5.08 (1H.m. PhCH₂ CHC), 5.33-6.01 (3H.m.β-lactams and PhCH), 7.52 (1H.s. Phthalide 3 proton), 6.95-8.05 (19H.m.aromatic protons and ##EQU9## 8.05-9.49 (2 × 1H.m. --CONH--*)

NH₂ OH Assay: 91%

Biochromatogram; Single zone R_(f) 0.87

EXAMPLE 46 D-α-[L-β-Phenyl-α-Ureidopropionamido]-PhenylacetamidoPenicillanic Acid

(R = Ph ; R¹ = PhCH₂ ; R³ = H ; R² = --NHCONH₂ ; M = H ; α¹ = L).

Prepared by method (Bi) from D-α-Ureido-β-phenyl propionic acid.

Yield: 15%

ν_(max) (KBr): 3360 (Br.), 1775, 1650, 1525, 1315, 1230 and 705cm⁻ ¹.

δ[(CD₃)₂ SO]: 1.44 (3H.s. gem methyl), 1.59 (3H.s. gem methyl), 2.89(2H.m. PhCH₂₋₋), 4.25 (1H.s. C3 proton), 4.68 (1H.m. PhCH₂ CH), 5.64(5H.m. β-lactams NHCONH₂ *, PhCH), 6.27 (1H.d. --NHCO--), 7.28 (10H.m.aromatic protons), 8.62 and 9.17 (2 × 1H.d. --CONH--*)

Biochromatogram: Single zone

EXAMPLE 47D-α-[D,L-α-(3-ETHYLUREIDO)-β-PHENYLPROPIONAMIDO]-PHENYLACETAMIDOPENICILLANICACID ##SPC24##

R³ = h, r² = nhconhch₂ ch₃, m = h, α¹ = d,l).

prepared by method (Bi) from D,L-(3-ethylureido)β-phenylpropionic acid.

Yield: 16% m.p. 174-6°C (dec.)

ν_(max) (KBr): 3380 (broad), 1774, 1637, 1540, 1299, 1218 and701cm.sup.⁻¹.

δ[(CD₃)₂ SO]; 0.96 (3H.t.--NHCH₂ CH₂), 1.44 (3H.s. gem methyl), 1.58(3H.s. gem methyl), 2.96 (4H.m. PhCH₂ CH<, --NHCH₂ CH₃), 4.27 (1H.s. C3proton), 4.59 (1H.m. PhCH₂ CH<), 5.38-6.17 (5H.m. β-lactams, PhCH<,--NHCONH--*), 7.32 (10H.m. PhCH<, PhCH₂ CH<), 8.58 (1H.m. --CONH--*),9.12 (1H.m.--CONH--*).

Hydroxylaminl Assay: 92.9%

Biochromatography: R_(f) = 0.71

EXAMPLE 48D-α-[D,L-β-(2-THIENYL)-α-UREIDOPROPIONAMIDO]PHENYLACETAMIDOPENICILLANICACID ##SPC25##

R³ = h--, r² = --nhconh₂, m = h, α¹ = d,l).

prepared by method (Bi) from D,L-β-(2-thienyl)α-ureidopropionic acid.

Yield: 25.9%

ν.sub. max (KBr): 3355 (br.), 1773, 1648, 1537, 1307, 1226 and701cm.sup.⁻¹.

δ[(CD₃)₂ SO]: 1.42 (3H.s. gem methyl), 1.56 (3H.s. gem methyl), 3.10(2H.m. --CH₂ CH<), 4.21 (1H.s. C3proton), 4.50 (1H.m. -CH₂ CH<), 5.58(5H.m. β-lactams, -NHCONH₂ *, PhCH<), 6.28 (1H.m. --NHCONH₂ *), 6.89(2H.m. thienyl 3- and 4- protons), 7.35 (6H.s. phenyl aromatics andthienyl 5- proton), 8.67 (1H.m. --CONH--*), 9.15 (1H.m. --CONH--*).

Hydroxylamine Assay: 70.6%

Biochromatography: R_(f) = 0.41

EXAMPLE 49 POTASSIUMD-α-[D-α-(3-ETHYLUREIDO)-p-PHENYL-PROPIONAMIDO]-PHENYLACETAMIDOPENICILIANATE

(R = Ph--, R¹ = PhCH₂ --, R³ = H--, R² = --NHCONHCH₂ CH₃, N = K, α¹ =D).

Prepared by method (Bi) from D-α-(3-ethylureido)-β-phenylpropionic acid.

Yield: 65.3%

ν _(max) (KBr): 3350(br), 1774, 1630, 1540, 1225, 732 and 702cm.sup.⁻¹.

δ [(CD₃)₂ SO]: 0.95 (3H.t.J = 7Hz --NHCH₂ CH₃), 1.45 (3H.s. gem-methyl),1.58 (3H.s. gem-methyl), 2.75-3.35 (4H.m. PhCH₂ CH, --NHCH₂ CH₃ (J =7Hz)), 4.28 (1H.s. C-3 proton), 4.61 (1H.m. PhCH₂ CH<), 5.4-6.3 (5H.m.β-lactams, PhCH: --NHCONH--*); 7.37 (10H.m. PhCH . PhCH₂ CH<), 8.53(1H.d. --CONH--*), 9.12 (1H.m. --CONH--*),

Hydroxylamine Assay: 85.4%

Biochromatography: R_(f) = 0.65

EXAMPLE 50D-α-[D-β-PHENYL-α-(3-n-PROPYLUREIDO)-PROPIONAMIDO]PHENYLACETAMIDOPENICILLANICACID ##SPC26##

R³ = H, R² = NHCONHCH₂ CH₂ CH₃, M = H, α¹ = D)

Prepared by method (Bi) from D-α-(3-n-propylureido)-propionic acid.

Yield: 70.3%

ν.sub. max (KBr); 3320 (br.), 1772, 1633, 1540, 1222, 731 and701cm.sup.⁻¹.

δ[(CD₃)₂ SO]: 0.81 (3H.t. --NHCH₂ CH₂ CH₃), 1.45 (3H.s. gem methyl),1.58 (3H.s. gem methyl), 2.7-3.3 (6H.m. PhCH₂ CH<, --NHCH₂ CH₂ CH₃),4.27 (1H.s. C-3 proton), 4.6 (1H.m. PhCH₂ CH<), 5.4-6.3 (5H.m.β-lactams, PhCH<, --NHCONH--*), 7.31 (10H.m. PhCH<, PhCH₂ CH<), 8.53(1H.m. --CONH--*), 9.10 (1H.m. --CONH--*).

Hydroxylamine Assay: 94.7%

Biochromatography; R_(f) = 0.73

EXAMPLE 51D-α-[D-β-PHENYL-α-(3-ISO-PROPYLUREIDO)-PROPIONAMIDO]-PHENYLACETAMIDOPENICILLANICACID ##SPC27##

R³ = h--, r² = --nhconhch(ch₃)₂ m = h, α¹ = d).

prepared by method (Bi) from D-α-phenyl-α-(3-iso-propylureido)-propionicacid.

Yield: 60%

ν.sub. max (KBr); 3363 (br), 1772, 1626, 1533, 1230, 1128, 729 and 701cm.sup.⁻¹.

δ[(CD₃)₂ SO]: 0.99 (6H.d. NHCH(CH₃)₂), 1.45 (3H.s. gem methyl), 1.58(3H.s. gem methyl), 2.92 (2H.m. PhCH₂ CH<), 3.3. (1H.m. --NHCH(CH₃)₂),4.28 (1H.s. C-3 proton), 4.6 (1H.m. PhCH₂ CH<), 5.4-6.3 (5H.m.β-lactams, PhCH<, --NHCONH--*), 7.23 (5H.s. PhCH₂ CH<), 7.39 (5H.m.PhCH<), 8.4-9.5 (2H.m. 2 × --CONH--*).

* Removable with D₂ O

Hydroxylamine Assay: 89.7%

Biochromatography: R_(f) = 0.72

EXAMPLE 52 POTASSIUMD-α-[D-α-(3-CYCLOHEXYLUREIDO)-β-PHENYLPROPIONAMIDO]-PHENYLACETAMIDOPENICILLANATE##SPC28##

M = k, α¹ = d.

prepared by method (Bi) from D-α-(3-cyclohexylureido)-β-phenylpropionicacid.

Yield: 57%

ν.sub. max (KBr): 3330 (br.), 1762, 1628, 1546, 1392, 1320 and701cm.sup.⁻¹.

δ[(CD₃)₂ SO]: 0.6-1.7 (10H.m. cyclohexyl methylenes) 1.44 (3H.s gemmethyl), 1.54 (3H.s. gem methyl), 2.75 - 3.4 (3H.m. PhCH₂ CH<, -NH-CH),3.92 (1H.d. C-3 proton), 4.3-4.7 (1H.m. PhCH₂ CH<), 5.2-5.6 (2H.m.β-lactams), 5.65-5.94 (1H.m. PhCH<), 6.1-6.5 (2H.m. ---NHCONH--*), 7.23(5H.s. PhCH₂ CH<), 7.37 (5H.m. PhCH<), 8.4-9.0 (2H.m. 2 x --CONH--*)

Hydroxylamine Assay: 85.9%

Biochromatography: R_(f) = 0.73

EXAMPLE 53 Potassium D-α-[DLα-(3-tert-butylureido)-β-phenylpropionamido]phenylacetamidopenicillanate

(R= Ph; R¹ =PhCH₂ --; R³ =H; R² = --NHCONHC(CH₃)₃ ; M=K; α¹ =D,L).Prepared by method (Bi) from D,L-α-(3-tert-butylureido)-β-phenylpropionic acid.

Yield: 40.2%.

ν.sub. max (KBr): 3360(br), 1774, 1645(br), 1540(br), 1456, 1214, 733and 702 cm.sup.⁻¹

δ[(CD₃)₂ SO]: 1.22 (9H.s. -NHC(CH₃)₃); 1.45(3H.s. gem dimethyl);1.58(3H.s. gem dimethyl); 2.89(2H.m. PhCH₂ CH<) 4.23 (1H.s. C-3 proton);4.53(1H.m. PhCH₂ CH<) 5.35-6.15(5H.m. β-lactams, PhCH < and--NHCONH--*); 7.30 (10H.m.PhCH < and PhCH₂ CH <); 8.51(1H.d. --CONH--*);9.12(1H d. --CONH--*).

Hydroxylamine Assay: 99.8%.

Biochromatography: 1 zone at Rf 0.72

EXAMPLE 54 D-β-[DL-β-(p-Hydroxyphenyl)-α-ureidopropionamido]phenylacetamidopenicillanic acid

R=Ph; R¹ =p-HO--PhCH₂ ; R³ =H; R² -- --NHCONH₂ ; M=H; α¹ =D,L. Preparedby method B from D,L-β-(p-hydroxyphenyl)-α-ureidopropionic acid.

Yield: 80%.

ν.sub. max (KBr): 3350(br), 1772, 1650, 1517, 1230, and 703 cm.sup.⁻¹

δ[(CD₃)₂ SO]: 1.44(3H.s. gem methyl); 1.57 (3H.s. gem methyl) ˜2.8(2H.m. --CH₂ CH<); 4.29 (1H.s. C-3 proton) ˜4.5 (1H.m -CH₂ CH<);5.35-5.87 (5H.m. β-lactams, PhCH< and >NHCONH₂ *); 6.27 (1H.d. -NHCONH₂*); 6.67 (2H.d. o-protons in p-HO-Ph ring); 6.99 (2H.d. m-protons inp-HO--Ph ring); 7.30(5H. broad s. PhCH ); 8.47 (1H.m. --CONH--*) 9.12(1H.m. --CONH--*)

Hydroxylamine Assay: 79.4%.

Biochromatography: 1Zone at Rf 0.27.

EXAMPLE 55

D-α-[D,L,β-(m-Hydroxyphenyl)-α-ureidopropionamido]-phenylacetamidopenicillanic acid.

(R=Ph, R¹ = m-HO--PhCH₂ --, R³ =H, R² = --NHCONH₂, M=H, α¹ =D,L ).Prepared by method B from D,L-β-(m-hydroxyphenyl)-α-ureidopropionicacid.

Yield: 46%.

ν_(max) (KBr): 3360(br), 1775, 1625, 1531, 1236, 1164, and 703 cm⁻ ¹

δ[(CD₃)₂ SO]: 1.45 (3H.s. gem methyl); 1.58 (3H.s. gem methyl);˜2.8(2H.m. --CH₂ CH<); 4.32 (1H.s. C-3 proton) 4.55 (1H.m. --CH₂ CH<);5.40-5.90 (5H.m. β-lactams, PhCH< and --NHCONH₂ *); 6.35 (1H.d.--NHCONH₂ *) 6.55-7.15 (4H.m. m-HO--Ph-- aromatics); 7.32 (5H. broad s.PhCH<); 8.52 (1H. d. --CONH--*); 9.16 (1H.m. --CONH--*)

Hydroxylamine Assay: 93.7%.

Biochromatography: 1Zone at Rf 0.39

EXAMPLE 56D-α-[D-β-Phenyl-α-ureidopropionamido]-(2-thienyl)-acetamidopenicillanicacid. ##SPC29##

R¹ =phCH₂ --, R³ =H, R² = --NHCONH₂, M=H, α¹ =D). Prepared by method(Bviii) with D-β-phenyl-α-ureidopropionic acid.

Yield: 44.8%.

ν_(max) (Nujol); 3310(br), 1785, 1668, 1539, 1233 and 701 cm⁻ ¹

δ[(CD₃)₂ SO]: 1.47(3H.s. gem methyl); 1.60(3H.s. gem methyl;˜ 3(2H.m.PhCH₂ CH<); 3.9-4.4(1H.s. PhCH₂ CH<) 4.26(1H.s. C-3 proton); 5.3-6.6(6H.m. β-lactams, ThCH<, and -NHCONH₂ *); 7.28(8H. m. PhCH₂ CH< andThCH<); 7.7-9.5 (2H.m. 2x --CONH--*)

Hydroxylamine Assay: 78.2%.

Biochromatography: One zone at Rf 0.62 (plus faint zone at Rf 0.27 dueto starting amino penicillin.)

EXAMPLE 57 D-α-[D-β-Phenyl-α-ureidopropionamido]-valeramidopenicillanicacid.

(R= CH₃ (CH₂)₂ --, R¹ = PhCH₂ --, R³ =H, R² =)NHCONH₂, M=H, α¹ =D).Prepared by method (Bvii) from D-β -phenyl-α-ureidopropionic acid.

Yield: 30%. ν_(max) (KBr): 3340(br), 1774, 1650, 1530, 1231, and 703cm⁻¹

δ[(CD₃)₂ SO]: 0.92 (3 H.m.CH₃ (CH₂)₂ --); 1.1-1.7 (1OH. m. gemdimethyls,CH₃ (CH₂)₂ --); 2.85-3.0 (2H.m. PhCH₂ CH<); 4.3-4.7 (2H.m. CH₃ (CH₂)₂CH<, PhCH₂ CH<); 4.34 (1H.s. C-3 proton); 5.47-5.8 (4H.m. β-lactams,--NHCONH₂ *); 6.30 (1H.d. --NHCONH₂ *); 7.27 (5H.s. aromatics); 8.12(1H.d. --CONH--*); 8.81 (1H.m. --CONH--*)

Hydroxylamine Assay: 78%.

Biochromatography: 1 zone at Rf 0.59.

EXAMPLE 58D-α-[D-β)Phenyl-α-ureidopropionamido]-cyclopropylacetamidopenicillanicacid. ##SPC30##

R¹ =PhCH₂ -, R³ =H, R² = -NHCONH₂, M=H, α¹ =D), Prepared by method (Bvi)from D-β-phenyl-α-ureidopropionic acid.

Yield: 50%.

ν_(max) (KBr): 3345(br), 1772, 1645,(br), 1527, 1230 and 703 cm⁻ ¹

δ[(CD₃)₂ SO]: 0.25-1.25H.m.cyclopropyl ring protons) 1.51 (3H.s. gemmethyl); 1.63 (3H.s. gem methyl); 2.92 (2H.m. PhCH₂ --); ##SPC31##

4.3 (1h.s. PhCH₂ CH<) 5.5-5.8 (4H.m. β-lactams and --COHN₂ *); 6.18(1H.d. --CONH--*) 7.2-7.4 (5H.m. aromatic protons); 8.18 and 8.93(2x1H.d. --CONH--*)

Hydroxylamine Assay: 95.6%.

Biochromatography: 1 zone at Rf 0.37

EXAMPLE 59D-α-[D-β-PHENYL-α-UREIDOPROPIONAMIDO]-β-PHENYLPROPIONAMIDOPENICILLANICACID

(R = R¹ = PhCH₂ -, R³ = H, R² = -NHCONH₂, M=H, α¹ = D).

Prepared by method (Bix) from D-p-phenyl-α-ureidopropionic acid.

Yield: 40%

ν_(max) (KBr): 3322(br), 1725, 1638, 1534, 1302, 1231, 702cm⁻ ¹

δ[(CD)₃ SO]: 1.51 (3H.s. gem-methyl), 1.65 (3H.s. gem-methyl), 3.0(4H.m. PhCH₂, PhCH₂), 4.32 (1H.s. C-3 proton), 4.4 and 4.8 (2 × 1H.m.PhCH₂ CH< ), 5.52 (2H.m. β-lactams), 6.1 (1H.m. --CONH--), 7.1-7.4(10H.m. aromatic protons), 8.20 and 8.78 (2 × 1H.m. --CONH*--)

NH₂ OH Assay: 87%

Biochromatography: Single zone R_(f) 0.40

EXAMPLE 60 D-α-[D-α-UREIDO-n-HEPTANAMIDO]-PHENYLACETAMIDO PENICILLANICACID (R = Ph; R¹ = CH₃ (CH₂)₅ ; R³ = H; R² = -NHCONH₂ ; M = H; α¹ = D).

Prepared by method (Bi), from D-Ureidoheptanoic acid.

Yield: 45%

ν_(max) (KB ): 3380(br), 1763, 1650, 1600, 1538, 1401, 1323, 1234,699cm⁻ ¹.

δ[(CD₃)₂ SO] : 0.87 (3H.m. CH₃ (CH₂)₅), 1.0-2.0 (10H.m. CH₃ (CH₂)₅),1.47 (3H.s. gem-methyl), 1.56 (3H.s. gem-methyl), 3.97 (1.H.s. C-3proton), 3.4-3.8 (1H.m. (CH₂)₅ CH<), 5.3-5.9 (5H.m. β-lactams, PhCH<,--CONH₂ *), 7.2-7.6 (5H.m. aromaticprotons), 6.6, 8.7 and 8.9 (3 × 1H.m.--CONH*--)

NH₂ OH Assay: 37%

Biochromatography: Single zone R_(f) 0.67

EXAMPLE 61 D-α-[D-α-UREIDO-n-HEXANAMIDO]-PHENYLACETAMIDO PENICILLANICACID

(R = Ph, R¹ = CH₃ (CH₂)₃, R³ = H, R² = -NHCONH₂, M = H, α¹ = D.)

Prepared by method Bi) using D-α-ureidohexanoic acid.

Yield: 60%

ν_(max) (KBν): 3340, 1772, 1640, 1312, 1234, 700cm⁻ ¹.

δ[(CD₃)₂ SO] : 0.84 (3H.m. CH₃ (CH₂)₃, 1.0-1.8 (6H.m. CH₃ (CH₂)₃, 1.42(3H.s. gem-methyl), 1.54 (3H.s. gem-methyl), 4.23 (1H.s. C-3 proton),4.23 (1H.m. CH₃ (CH₂)₃ CH<), 5.4-5.9 (5H.M. β-lactams, Ph.CH<and CONH₂*), 6.25 (1H.m. --CONH*--), 7.40 (5H.m. aromatic protons), 8.49 and 9.08(2 × 1H.d. --CONH* --)

Hydroxylamine Assay: 73%

Biochromatography: R_(f) = 0.59

EXAMPLE 62D-α-[D-β-(1,4-Cyclohexadienyl)-α-Ureidopropionamido]-(p-Hydroxyphenyl)AcetamidopenicillanicAcid

(R =_(p) --HO--Ph), ##SPC32##

R³ = h, r² = --nhconh₂, m = h, α¹ = d)

prepared by method (Bii) usingD-β-(1,4-cyclohexadienyl)-α-ureidopropionic acid

Yield: 50%

ν_(max) (KBν): 3330(br), 1770, 1640, 1510, 1223, 961, and 840cm.sup.⁻¹.

δ[(CD₃)₂ SO] : 1.45 (3H.s. gem-methyl); 1.58 (3H.s. gem-methyl);##SPC33##

2.61 (4H. broad s. cyclohexadiene methylenes)); 4.24 (1H.s. C-3 proton);##SPC34##

5.35-5.8 (8H.m. β-lactams, ##SPC35##

cyclohexadiene methines, --NHCONH₂ *); 5.9-6.2 (1H.m. --NHCONH₂ *);##SPC36## ##SPC37##

8.3 (1h.m. --CONH--*); 8.9 (1H.m. --CONH--*).

Hydroxylamine Assay: 98.2%

Biochromatography: R_(f) =0.41

EXAMPLE 63D-α-[D-β-(1,4-Cyclohexadienyl)-α-(3-Ethylureido)-Propionamido]-(p-Hydroxyphenyl)-Acetamido-PenicillanicAcid ##SPC38##

R³ = h--, r² = --nhconhch₂ ch₃, m = h, α¹ = d).

prepared by method (Bii) fromD-β-(1,4-cyclohexadienyl)-α-(3-ethylureido)-propionic acid.

Yield: 20%

ν_(max) (KBr): 3350 (br), 1760, 1510, 1371, 1258, 1220 and 781cm.sup.⁻¹.

δ[(CD₃)₂ SO]: 1.1-1.7 (9H.m. gem dimethyls, --NHCH₂ CH₃) ##SPC39##

2.45-2.75 (4h.m. cyclohexadiene methylenes), 3.9-4.5(44.m. NHCH₂ CH₃,C-3 proton and ##SPC40##

5.0-5.8 (8H.m. β-lactams, ##SPC41##

cyclohexadiene methines, --NH*CONH--*), 6.6-7.6 (4H.m. aromatics),8.3-9.2 (2H.m. 2 x --CONH--*).

Hydroxylamine Assay: 88%

Biochromatography: R_(f) = 0.44 (plus 2 minor zones)

EXAMPLE 64D-α-[D-α-Acetamido-n-Hexanamido]-(p-Hydroxyphenyl)-AcetamidopenicillanicAcid ##SPC42##

R¹ = ch₃ (ch₂)₃ --, r³ = h--, r² = --nhcoch₃, m = h, α¹ = d).

prepared by method (Bii) from D-α-acetamido-n-hexanoic acid.

Yield: 30%

ν_(max) (KBr): 3310(br.), 1770, 1645, 1510, 1374, 1210, 1173cm.sup.⁻¹.

δ[(CD₃)₂ SO]: 0.8-1.8 (9H.m. CH₃ (CH₂)₃ --), 1.43 (3H.s. gem methyl),1.54 (3H.s. gem methyl), 1.92 (3H.d. COCH₃) 4.28 (1H.s. C3 proton), 4.38(1H.m. CH₂ CH<), 5.4-5.8 (3H.m. β-lactams and PhCh<), 6.22 (1H.d.--CONH--*), 7.0-7.6(4H.m. aromatic protons), 8.25 and 9.3 (2 × 1H.m.--CONH--*)

Hydroxylamine Assay: 124.6%

Biochromatography: R_(f) = 0.35 (plus small zone due toamino-penicillin).

We claim:
 1. A penicillin of formula (I): ##SPC43##wherein R is phenyl,hydroxyphenyl, halophenyl, nitrophenyl, alkoxyphenyl having 1-3 carbonatoms in the alkoxy part, aminophenyl, 2- or 3-thienyl, cycloalkylhaving 3-7 carbon atoms in the cyclo part, cycloalkenyl having 5-7carbon atoms in the cyclo part, or alkyl having 1-4 carbon atoms; R¹ isacetoxy; R² is ##SPC44## in which R⁵ is amino, mono- or di-alkylaminohaving 1-4 carbon atoms in the alkyl part, cyclohexylamino, hydrogen,alkyl of 1-4 carbon atoms or phenyl, and R⁶ is amino, mono- ordi-alkylamino having 1-4 carbon atoms in the alkyl part orcyclohexylamino; R³ is hydrogen or alkyl of 1-3 carbon atoms; or apharmaceutically acceptable salt or a pharmaceutically acceptablehydrolyzable ester which converts to the free acid form in vivo.
 2. Apenicillin of claim 1 wherein the carbon atom attached to R is in theD-configuration.
 3. A penicillin of claim 1 wherein the carbon atomattached to R² is in the D-configuration.
 4. A penicillin of claim 1wherein R is phenyl, 4-hydroxyphenyl or 3-thienyl.
 5. A penicillin ofclaim 1 wherein R³ is hydrogen.
 6. A penicillin of claim 1 wherein, whenR² is (II), R⁵ is amino or hydrogen or, when R² is (III), R⁶ is amino.7. A penicillin of claim 1 wherein the ester is phthalidyl,5,6-dimethoxyphthalidyl, pivaloyloxymethyl or acetoxymethyl.
 8. Apenicillin of claim 1 wherein the salt is a base addition or acidaddition salt.